Cellulolytic Bacterial Isolates Research Articles

Microbial Protein Synthesis by Cellulolytic Bacterial Isolates from Feces of Indonesian Endemic Herbivores

Cellulolytic bacterial play an important role in the synthesis of microbial protein in the small intestine as high-quality protein. This research aimed to observe the growth patterns of cellulolytic bacterial isolates from the feces of endemic herbivorous animals in Indonesia, and to evaluate the effect of the addition of cellulolytic bacterial isolates on the synthesis of microbial proteins in vitro. The study used a completely randomized design with four treatments and five replications. The treatments included T0 = Concentrate mix 40%: elephant grass 30%: palm leaves 30% (control), T1 = T0 + 105 CFU mL-1 cellulolytic bacterial isolate, T2 = T0 + 106 CFU mL-1 cellulolytic bacterial isolate, and T3 = T0 + 107 CFU mL-1 cellulolytic bacterial isolate. Variables measured were the growth pattern of bacterial isolates, bacterial population at the optimal growth point, and microbial protein synthesis. The results showed growth pattern of cellulolytic bacterial isolates from Muntjac, Deer and Anoa feces have similar pattern, and was different from the growth pattern of bacterial isolates from bison feces. The optimal growth point of cellulolytic bacterial isolates from the feces of Muntjac, Deer, and Anoa at the 8th hour of incubation with bacterial populations of 1.3 x 109 CFU mL-1, 1.7 x 109 CFU mL-1, and 1.4 x 109 CFU mL-1. The growth pattern and optimal point of cellulolytic bacterial isolates from the feces of Bison at the 12th hour after incubation with bacterial populations of 1,1 x 1010 CFU mL-1. The addition of cellulolytic bacterial isolates did not affect microbial protein synthesis.

A Comparative Study of Different Staining Techniques for Cellulase Activity on CMC (Carboxy Methyl Cellulose) Agar

The present experiment was conducted at Department of Plant Pathology, College of Agriculture, Navsari Agricultural University, WaghaiduringMarch-October, 2021. cellulolytic bacterial isolates CD11, CD17, CD19, CD22 and CD35 were screened to determine the staining efficiency of various stains for cellulase activity on CMC(Carboxy Methyl Cellulose) agar and it was observed that all the isolates conferred clear zone around the colonies through different staining dyes and bacterial isolate CD35 showed the highest cellulolytic index in all the dyes. The cellulolytic index of CD35 was highest with gram’s iodine (3.34) next in order of coomassie brilliant blue (2.96), safranin (2.55) and congo red (2.15).Significantly, higher cellulase activity was recorded in CD35 (0.169 Uml-1) at 24 hrs after inoculation It was followed by CD17 (0.124 Uml-1), CD19 (0.101 Uml-1) and CD11 (0.081 Uml-1), while it was lowest in CD22 (0.052 Uml-1). Maximum cellulase activity was recorded at 72 hrs after inoculation by all the selected isolates except CD22 isolate, which was maximum 96 hrs after inoculation. CD35 gave significantly maximum cellulase activity (0.822 Uml-1) at 72 hrs after inoculation. Next in order to cellulase activity was CD17 (0.477 Uml-1) which was at par with CD19 (0.471 Uml-1) followed by CD11 (0.292 Uml-1) while, it was lowest in CD22 (0.199 Uml-1).Cellulolytic bacteria CD35 was identified as Bacillus subtilis by morphological, biochemical and molecular methods and submitted to NCBI GenBank database with accession numbers MW715021.

Undesirable Microbial Growth from The Biodegradation Process of Nypa Palm Fronds Suplemented with Cellulolytic Bacteria Culture Isolated from Nypa Palm Worms (Namalycastis rhodochorde)

Biodegradation of nipa palm fronds using cellulolytic bacteria on an in-vitro scale was conducted to produce feed for cultivating palm worms (Namalycastis rhodochorde). The spontaneous degradation process is thought to trigger the growth of other microbes from raw materials from unsterilized nypa palm fronds. This research aims to determine the presence of different microbes that participate in the degradation process of palm fronds supplemented with cellulolytic bacterial isolates. The biodegradation process was conducted using dry palm fronds that were not sterilized, without pressurized heat, for 30 days in a sterile container. Cellulolytic bacterial isolates were added to facilitate the process. The research results revealed three characteristics of bacterial colonies with two cell shapes: bacilli and spirals bacterial isolates. The study also identified the presence of protozoa that exhibited morphological similarities to the Ciliophora group. This research provides valuable reports for further studies on the biodegradation process of palm fronds, specifically about ensuring feed safety for cultivating nypa palm worms.

Grasshopper (Oxya chinensis) Gut Bacteria and their Cellulolytic Activity

Lignocellulose is a core component of plant biomass and the most abundant carbohydrate polymer in nature. It is cheap and renewable and has several potential applications; however, it remains underutilized because of its recalcitrance to degradation. Cellulolytic microbes have been found in the gut of herbivorous insects, such as grasshoppers. This study aimed to isolate lignocellulolytic bacteria from the gut of grasshoppers (Oxya chinensis) and determine their diversity and potential biomass-degrading activity. A total of 27 culturable isolates were obtained from the grasshopper foregut, midgut, and hindgut. The bacteria hydrolyzed cellulose and lignin, as indicated by a cellulolytic index of 0.12–1.23 and ligninolytic index of 0.1–1.47. Five potential cellulolytic bacterial isolates were selected. Based on 16S rRNA sequencing, the isolates were identified as Bacillus wiedmannii (foregut), Bacillus marcorestinctum, Bacillus halotolerans (midgut), Paenibacillus zanthoxyli, and Bacillus hominis (hindgut). The highest specific cellulolytic activity (0.0068 U/mg) was detected in B. wiedmannii (OCF2), which could be exploited as a potential source of cellulases.

The Impact of Peatland Fires on Cellulolytic Microbial Populations in Several Landuses

Studies on the impact of peatland fires on agricultural land, especially oil palm, and rubber, are still limited. This study aims to calculate the total population of cellulolytic bacteria and determine the activity of cellulolytic enzymes qualitatively in each microbial isolate found and to determine the level of peatland degradation after the fire. The research was conducted by survey in the field. The research location is in the long jungle village of Tambang Subdistrict, Kampar Regency. At the same time, the laboratory analysis was carried out in the Pathology, Entomology, and Microbiology Laboratory, Faculty of Agriculture and Animal Husbandry, Sultan Syarif Kasim Riau University. Determination of sample points using the purposive sampling method, which is to choose a location per the study's objectives at 4 locations. The first location is oil palm land that is not burnt, the second is burning oil palm land, the third is unburned rubber land, and the fourth is burned rubber land. Each location was repeated three times to obtain 12 sample points for observation. Furthermore, at each observation sample point, two soil depths were observed, 0-15 cm and 15-30 cm. The total population of cellulolytic bacteria in oil palm and rubber plantations that were burned and not burned with different soil depths ranged from 5.3 x 109 cfu / g soil to 14.0 x 109 cfu / g soil. The clear zone ratio (Z / K) of 8 cellulolytic bacterial isolates observed was high criteria, namely> 1.76 (100%). The highest Z / K ratio was produced by bacterial isolates originating from burnt rubber plantations at a soil depth of 0-15 cm, namely 5.25.

Isolation and Molecular Identification of Cellulase producing bacteria from soil and litter in Veerakkal forest area, The Western Ghats, India

The cellulolytic bacterial strains were screened from both soil and litter samples in different study sites of Veerakkal forest, Manar beat, The Western Ghats, India. In the current investigation, out of 64 and 48 differentiated total purified isolates, 53 soil isolates and 34 litter isolates were screened based on the CMC method. The percent wise calculation indicated that 82 % were cellulase positive while 18 % were negative in soil isolates. Likewise, 70 % of them were positive while 30 % were negative cellulase strains in litter isolates. The average hydrolysis capacity (HC) values ranged between 1.67 to 3.25 cm. The highly potential cellulolytic bacterial isolates viz. Bacillus thuringiensis, Bacillus licheniformis, Bacillus cereus, Bacillus sporothermodurans, Bacillus circulans, Bacillus albus, Bacillus amyloliquefaciens, Lysinibacillus sphaericus and Bacillus mobilis were identified from soil. Likewise, the litter cellulolytic bacterial species such as Bacillus sp., Lysinibacillus macroides, Bacillus paramycoides, Solibacillus silvestris, Pseudomonas stutzeri, Bacillus toyonensis, Bacillus subtilis, Bacillus pacificus, Bacillus flexus, Bacillus paranthracis and Pseudomonas putida were identified. The cellulolytic bacterial population was higher during the monsoon because of the presence of higher moisture content in leaf litter. However, the cellulolytic bacterial population of soil was higher during the pre-monsoon season because of the continuous breakdown of litter which inhaled more cellulolytic strains in the soil. Finally, it was concluded that corresponding with seasonal changes, the cellulolytic bacterial community also changed and it plays a significant role in the mineralization process.

Cellulolytic Bacteria as Decomposers in Dry Empty Fruit Bunches Composting

Empty fruit bunches (EFB) contain macro and micronutrients, so they can be used as compost. EFB takes a long time to be composted because it has high cellulose. Used cellulolytic bacteria decomposers are a solution to boost EFB composting. The goal of this research is to get cellulolytic bacteria with a high potential for accelerating composting and producing high-quality compost. The study used a completely randomized design with several types of cellulolytic bacteria and was repeated three times. Data is presented in qualitative and quantitative formats. The qualitative data were analyzed descriptively, the quantitative data were analyzed using analysis of variance, and the Honestly Significant Difference (HSD) test was run at a 5% significance level. Temperature, volume shrinkage, color, smell, C-organic, N-total, C/N ratio, and pH parameters of the compost were analyzed. The results showed that the application of cellulolytic bacterial isolates could improve the quality of OPEFB compost (in accordance with SNI Compost Quality Standards: 19-7030-2004) and speed up composting time. Consortium I was the best treatment with reduced compost volume up to 74.53%, decreased C-organic content up to 23.32%, increased total N content up to 1.88% and decreased C/N ratio up to 12.44.

The effect of lignocellulolytic bacteria consortium on composting empty oil palm fruit bunches

Oil palm empty fruit bunches (OPEFB) are waste produced by palm oil mills from processing fresh fruit bunches into oil. For every 1 tonne of fresh fruit bunch (FFB) processing, 21–23% EFB will be produced. EFB can be used as compost. OPEFB compost has considerable potential because it contains complex nutrients (macro and micro). The OPEFB composting process has problems because of the lignin content which causes the decomposition process to take a long time so that microorganisms are needed that can accelerate the OPEFB decomposition rate. This study aimed to test the ability of lignolytic bacteria from larvae of Oryctes rhinoceros L. plus cellulolytic bacteria from organic waste that have the potential to decompose OPEFB. This research was conducted experimentally using a non-factorial Completely Randomized Design (CRD) consisting of eight treatments. The results showed that the administration of cellulolytic bacterial isolates had a significant effect on water content, compost weight loss, pH, C-Organic, C/N ratio and P nutrient content, but had no significant effect on N and K nutrient levels. Treatment of lignocellulolytic bacteria ORL9 and ORL6 and a consortium of cellulolytic bacteria showed good compost quality, which could be seen from the parameters of weight loss of compost, pH, N nutrient content and reduced C/N ratio.

Isolasi dan Seleksi Bakteri Kandidat Selulolitik dari Proses Pembuatan Pupuk Organik pada Pengolahan Limbah Peternakan

Cattle farming is one of the livestock business commodities in Indonesia. In the production process, cattle farms produce waste in the form of feces and leftover feed which can be processed in such a way through a decomposition process into organic fertilizer. The rate of the decomposition process is influenced by the existing decomposer microbes. One of the microbes needed in the decomposition process is cellulolytic bacteria. The research was conducted to obtain potential cellulolytic bacterial isolates from the compost which can then be used as a starter in the decomposition process. This research was conducted experimentally with the stages of research including isolation of bacteria from decomposers, the isolates obtained were then reviewed on Nutrient Agar media to produce uniform colonies. Furthermore, screening was carried out to obtain cellulolytic bacteria using NA media added with 1% CMC (Carboxymethyl Cellulose). Based on the results of the study, obtained 7 bacterial isolates that have various cellulolytic indices. The best cellulolytic index was produced by isolates NC-12, NC-17, and NC-16.

MOLECULAR DIVERSITY AMONG CELLULOLYTIC BACTERIAL CULTURES ISOLATED FROM RURAL AND URBAN WASTES

A total of 34 samples of decomposing rural and urban wastes were collected from different partsof Western Maharashtra from which 64 bacteria, 49 fungi and 63 actinomycetes strains were isolated. Theseisolates were screened for cellulase activity and the efficient cellulolytic microorganisms were identified onthe basis of morphological, cultural and biochemical characters. Among cellulolytic bacterial isolates, basedon cultural, morphological and biochemical characteristics, 11 isolates were identified as Bacillus spp., 3 asPseudomonas spp., while the remaining three belonged to Cellulomonas, Staphylococcus and Micrococcusgenus. The Bacillus isolates were further studied for their molecular diversity within group. The PCRamplification products of the 12 isolates of Bacillus spp. including standard Bacillus strain with 16 randomprimers produced 343 bands, out of which 182 bands were polymorphic and 158 bands were unique.Majority of the primers showed 100% polymorphism. The average polymorphism observed was 99.13%. Thegenetic similarity index based on the pooled data of RAPD profiles from all the 16 primers ranged from 0.11to 0.76 among Bacillus isolates. The dendrogram constructed from the pooled data showed four majorgroups showing wide genetic variability between the isolates. There was wide genetic variability in Bacillusspp. at genus level from the same ecological niche.

Screening of Cellulolytic Bacteria from Biological Education and Research Forest Floor Andalas University, Indonesia.

<b>Background and Objective:</b> Organic waste dump is a problem that needs to be solved, one of which is by using microbe. Cellulolytic bacteria's ability to produce cellulase enzymes that can hydrolyze cellulose. Cellulose is the major component of the plant cell walls that are difficult to endure degradation naturally. This study aimed to find cellulolytic bacterial isolates on Biological Education and Research Forest floor Andalas University and characterize the cellulolytic bacteria found. Material and Methods: CMC (Carboxymethyl Cellulose) medium is used as a screening for bacteria isolate and this study used the survey method and also conducted catalase, glucose and lactose test for characterizations. <b>Results:</b> We found 16 bacterial isolates on Biological Education and Research Forest floor where 9 isolates were in a shaded area and 7 isolates were in an unshaded area. There were 12 isolates from 16 isolates that have the positive cellulolytic ability with a variety of clear zone sizes, where there were 8 isolates with the large clear zone and 4 isolates producing very small clear zones. Characteristics of bacterial isolates with the large clear zone obtained were 2 gram-positive coccus isolates with positive catalase test, 3 gram-negative coccus isolates with positive glucose test and 3 gram-negative bacilli isolates with negative fructose test. <b>Conclusion:</b> We identified 2 potential isolates with a cellulolytic index value greater than 2, isolate UCB 4 with a cellulolytic index value of 3.5 and UCB 6 with a cellulolytic index value of 2.2.

Cellulolytic activity of gut bacteria isolated from the eri silkworm larvae, Samia ricini, (Lepidoptera: Saturniidae)

The eri silkworm, Samia ricini, is associated with symbiotic bacteria believed to be playing several physiological functions to the host. The larvae of S. ricini were subjected to isolation of gut bacteria using culture-dependent 16S rRNA gene amplification and sequencing; and qualitative and quantitative enzymatic assays. The results of qualitative cellulolytic assays indicated that 58% of the screened gut bacterial isolates had cellulolytic activity. The cellulolytic bacterial isolates belonged to Bacillus sp. (88%), Pseudomonas sp. (8%) and Enterococcus sp. (4%). Quantitative assays indicated that the highest β-glucosidase (13.42 U/mg), endoglucanase (3.60 U/mg) and exoglucanase (2.60 U/mg) activity were in isolates ERI011, ERI009 and ERI097, respectively. Our results further indicate that significant activities of endogenous cellulases are present in the gut fluids of S. ricini. However, a comparative analysis of the cellulolytic activity showed that the endoglucanase activity of bacterial origin was 13 and 18-fold higher in the foregut and midgut, respectively, than that in gut fluids. Similarly, the β-glucosidase activity was 20 and ninefold higher compared with gut fluids from the same compartments. The higher cellulolytic activity of bacterial origin compared with that of insect origin suggests that more cellulose digestion may be aided by cellulases of bacterial origin. These cellulolytic gut bacterial isolates may be good sources for profiling novel genes and biomolecules for biotechnological application.

Produksi Enzim Selulase Kasar dari Isolat Bakteri B2S8 menggunakan Substrat Brangkasan Jagung dengan Perlakuan Konsentrasi Inokulum dan Komposisi Media yang berbeda

The purpose of this study was determined the media and concentration of cellulolytic bacterial isolates to produce high cellulase enzyme activity. Production of crude cellulase enzyme in media and concentration of different bacterial isolate used a factorial Randomized Block Design (RBD) which consist of two factors. The first factor was the media production of different cellulase enzyme consisting of 3 levels, namely media 1, 2 and 3. The second factor was the concentration of bacterial isolate consisting of 5 levels namely 1, 2, 3, 4 and 5%. This study used a B2S8 cellulolytic bacterial isolate that has the highest value of cellulase enzyme activity and the highest degradation rate of cellulose in previous studied and determined the ability of exoglucanase enzyme activity, endoglucanase enzyme and dissolved protein content produced from cellulolytic bacterial isolate. This study used Carboxymethyl Cellulose (CMC) for enzyme activity test and 1% corn stover as a substrate on the media to produce crude cellulase enzyme. The result showed that the highest cellulase enzyme activity in the third media and 5% cellulolytic bacterial inoculum concentration resulted in endoglucanase activity of 0.0332 IU/mL, exoglucanases enzyme activity of 0.0060 IU/mL, dissolved protein content in the amount of 0.5670 mg/mL, the specific endoglucanase activity of 0.0807 IU/mg and the specific activity of exoglucanase of 0.0123 IU/mg.
 Keywords: Cellulolytic bacteria, Cellulase enzymes, Enzyme activity, Corn stover

Isolasi Dan Karakterisasi Bakteri Selulolitik Biopori Sebagai Upaya Awal Percepatan Proses Pengomposan

The purpose of this study was to obtain cellulolytic bacterial isolates isolated from leaf litter in absorption holes biopori FMIPA Unesa and obtain the most optimal cellulolytic bacterial isolates in cellulose degradation. This research was an observational study and the data were analyzed descriptively. Stages of the study began with bacterial isolation, cellulolytic ability testing, and characterization of cellulolytic bacterial isolates. Bacterial isolation was carried out by the pour plate method, isolation was carried out by the streak plate method, cellulolytic testing was carried out using Carboxy Methyl Cellulose media which was given Congo red 1%. While the isolation characterization was done morphologically, physiologically, and biochemically. The results obtained 15 isolates of cellulolytic bacteria that were tested for their ability to degrade cellulose. Cellulolytic test results showed that 6 isolates, namely BS1, BS7, BS10, BS11, BS14, and BS15 had a cellulolytic index of 0.8, 0.8, 0.8, 0.8, 0.8, 0.8. and 1. Isolates BS15 is the most optimal isolate in cellulose degradation with characteristics of the punctiform colony, yellow pigmentation, entire edge, convex elevation, optical opaque and smooth surface, produces catalase enzymes, is non-motile and can ferment in glucose and starch but cannot ferment lactose and also a Gram-negative

Rice Straw Composting by Cellulolytic Bacteria Isolate and Its Application on Rice in Acid Sulfate Soils

High acidity in acid sulfate soils due to pyrite oxidation results in increased Al3+ and Fe2+ activity which inhibits the growth of rice plants. The application of organic matter (compost) is one of the technology to manage acid sulfate soil. This study aims to obtain cellulolytic bacterial isolates that are superior in composting and improving rice growth in acid sulfate soil. The experiment carries out in the laboratory and glasshouse of the Indonesian Swampland Agriculture Research Institute (ISARI), Banjarbaru, Indonesia on May-November 2017. The experimental to obtain cellulolytic bacteria and water content that can accelerate composting is arranged by factorial using a complete randomized design with three replication First factor were cellulolytic bacteria application (without application/control, BS 1.6, BS 1.9, BS 2.2 and BS 2.5), while the second factor was water content (50%, 100%, and 150%). The effect of compost application with cellulolytic bacterial to rice growth arranged by factorial completely randomized design with 3 replications. The first factor was cellulolytic bacteria application (without application/control, BS 1.6, BS 1.9, and BS 2.2), while the second factor was composting condition (muddy waterlogged and waterlogged 5 cm depth). The result showed that the ability of cellulolytic bacteria to reduce C/N straw was not different. Only differences in water content affect the reducing C/N ratio of straw. The average C/N ratio of straw compost made with 50%, 100%, and 150% water content is 35.59; 29.71, and 29.21. Application of compost made under muddy waterlogged and inoculated BS1.9 and BS2.2 can increase the number of tillers, while those inoculated BS1.6 and BS1.9 can increase the rice shoot dry weight of Inpara 2. The suggest that cellulolytic bacterial inoculation can improve the quality of compost so that the growth of rice is better.

Isolation and characterization of Soil Termites (Macrotermes gilvus) cellulolytic bacteria and activity determination of cellulase enzyme on newsprint substrates

The study aims to explore and identify cellulolytic bacteria of termites that have the potential to cellulose hydrolysis on newsprint into glucose. The cellulolytic bacteria are characterized by the formation of clear zones in the medium agar containing newsprint paper after 0.1% Congo red testing. The isolates that produce large clear zones are identified by the type of bacteria. Identification of bacteria morphologically in colonies, gram staining and biochemical tests. The activity of cellulose enzyme based on concentration of reducing sugar (glucose) produced from the cellulose hydrolysis on newsprint was tested by 3.5-dinitrosylicyl reagent using a UV-Vis spectrophotometer. The results showed that 6 cellulolytic bacterial isolates through the Congo red test, 2 isolates that have a large diameter of the clear zone are: Isolates C1 and D1 4.27 cm and 2.05 cm, respectively. Cellulase enzyme crudes of isolates C1 and D1 have an enzyme activity of 15.7 mU/mL and 2.33 mU/mL, protein concentration of 0.35 mg/mL and 1.18 mg/mL with specific activity of 45.17 mU/mg protein and 1.98 mU/mg protein. Based on the observation of the colony’s morphology in macroscopy, microscopy and biochemical test results, an isolate of C1 is the Provedencia sp and D1 isolate is the Bacillus sp.

OPTIMASI SUHU DAN pH TERHADAP AKTIVITAS ENZIM ENDOGLUKANASE MENGGUNAKAN RESPONSE SURFACE METHODOLOGY (RSM)

This study aims to determine the optimal temperature and pH for the growth of cellulolytic bacterial isolates to produce high cellulase enzyme activity. This study used one potential cellulolytic bacterial isolate B2S8 which had the highest cellulose degradation value and highest cellulase enzyme activity in previous studies. Carboxymethyl Cellulose (CMC) was used as a substrate on growth media and enzyme activity test. Optimization of temperature and pH on cellulase enzyme activity was done by Response Surface Methodology (RSM) by Central Composite Design (CCD). The results using Response Surface Methodology (RSM) showed that the highest endoglucanase enzyme activity was at 36.9 ºC and pH 6.9 was equal to 0.0269 IU/mL.
 Keywords: cellulolytic bacteria, endoglucanase enzyme, activity, Response Surface Methodology.

Skrining Bakteri Selulolitik Asal Saluran Pencernaan Rayap Untuk Mendekomposisi Tunggul Karet

The rubber tree stump is a source of white root fungus inoculums and as a source of infection that causes the death of rubber plant. The emergence of this disease is closely related to the cleanliness of the land such as leftover trees or stumps, shrubs and bushes which stacked or still in the ground. One of the symbionts organisms found in gut of termite and play a role in the decomposition of cellulose, are bacteria. Termite are social insects that are efficiently decompose lignocelluloses with the aid of their associated microbial symbionts located in termite gut. The purpose of this study was to obtain cellulolytic bacterial isolates are derived from gut of termite and getting an cellulolytic which showed the best ability in decomposing rubber tree stumps. The result termite samples from oil palm, rubber, and dry wood contain cellulolytic bacteria characterized by clear zones around bacterial colonies. Bacterial isolates showing the ability to degrade cellulose are 31 isolates and five isolates are safe for plants, animals, or mammals. The three bacterial isolates (NK 4, NS 4, and NS 5) used in the test on rubber tree stumps able to decompose rubber tree stump. Bacterial isolates were each obtained NK 4 13.52%, NS 4 18.40% and NS 5 17.88%.Keywords: bacteria cellulolytic, rubber tree stumps, termite

KINETIKA REAKSI ENZIMATIS EKSTRAK KASAR ENZIM SELULASE BAKTERI SELULOLITIK HASIL ISOLASI DARI BEKATUL

<p>Bran rice is a by-product of rice into rice milling process, the cellulose content of 40-60%, so the potential as a carbon source for the growth of microorganisms such as bacteria to produce enzymes particularly cellulolytic bacteria. The purpose of the study was to determine the diversity of the characters from the cellulolytic bacterial isolates and optimum conditions enzyme (cellulase enzymes rough) so that they can hydrolyze the cellulose to glucose with either rice bran. The characterization includes the determination of pH, temperature and time of optimum crude extract of bacterial cellulolytic enzyme cellulase, determination of Vmax and Km and molecular mass determination of cellulase.</p><p>Research methods include making media, regeneration of isolates, bacterial growth curve manufacturing, production of cellulase enzymes from bacterial cellulolytic rough at the optimum conditions, the kinetics of enzymatic reaction: substrate concentration factor of the reaction rate (with variation of the concentration of 0.50%, 0.75%, 1 , 00%, 1.25% and 1.50% (w / v)) followed by calculating the Vmax and Km.</p><p>The results showed that the enzyme cellulase of cellulolytic bacteria isolated from rice bran result that has optimum conditions at pH 7.5, temperature 50 ° C, 40 min incubation time to produce Vmax 0.0086 units / mL and Km 1.694%.</p>

Isolation, Screening, and Optimization of the Fermentation Conditions of Highly Cellulolytic Bacteria from the Hindgut of Holotrichia parallela Larvae (Coleoptera: Scarabaeidae)

From the hindgut contents of Holotrichia parallela, 93 cellulolytic bacterial isolates were isolated after enrichment in carboxymethyl cellulose medium. Among these isolates, a novel bacterium, designated HP207, with the highest endoglucanase productivity was selected for further study. This bacterium was identified as Pseudomonas sp. based on the results of the 16S ribosomal DNA analysis, morphological characteristics, and biochemical properties. The production of the endoglucanase was optimized by varying various physical culture conditions using a submerged fermentation method. Under the optimized fermentation conditions, the maximum endoglucanase activity of 1.432 U mL(-1) in bacterial cultures was obtained, higher than those of the most widely studied bacteria and fungi, which are the attractive candidates for the commercial producer of cellulase. And the crude endoglucanase enzyme was also highly thermostable; approximately 55% of the original activity was maintained after pretreatment at 70 °C for 1 h. Thus, from the present study, the bacterium can be added up to the database of cellulolytic bacteria.